支气管哮喘大鼠淋巴液中淋巴细胞线粒体跨膜电位变化及地塞米松对其干预的影响
【摘要】 目的 探讨哮喘大鼠淋巴液中淋巴细胞线粒体跨膜电位、Fas/FasL变化以及地塞米松干预的影响。方法 建立哮喘模型,收集激发后2、6、12、24、48 h淋巴液、血液、肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中淋巴细胞,流式细胞仪(FCM)检测淋巴细胞线粒体的跨膜电位(△·ψm),细胞免疫组化(SABC法)检测淋巴细胞表面Fas/FasL蛋白的表达。结果哮喘组淋巴液、血液、BALF淋巴细胞线粒体跨膜电位均较对照组和治疗组峰值升高(P<0.05),而对照组和治疗组之间则无显著性差异(P>0.05);与对照组、治疗组相比较,哮喘组淋巴液和BALF中淋巴细胞表面Fas、FasL蛋白光密度明显降低(P<0.001),而治疗组和对照组比较无显著性差异(P>0.05)。结论 哮喘大鼠淋巴液、血液、BALF中均存在明显的淋巴细胞早期凋亡障碍和Fas/FasL表达降低,尤其是哮喘淋巴液中淋巴细胞凋亡障碍和Fas/FasL降低程度较其血液、BALF更为明显;地塞米松可能是通过提高哮喘Fas/FasL蛋白的表达来加速淋巴细胞凋亡而发挥治疗作用。
【关键词】 哮喘;淋巴液;线粒体跨膜电位(△·ψm);Fas/FasL;凋亡
Lymphocyte mitochondrial membrane potential in the lymph of bronchial asthmaticrat and the effect of dexamethasone on it
XING Chaopin1 FENG Xuebin1 SONG Xiaodong2 et al医.学全.在.线网站www.lindalemus.com
1 Depertment of Pediatric,Binzhou Medical University,Binzhou 256603;2 Experimental Conter of Binzhou Medical University
【Abstract】 Objective To observe the levels of lymphocyte mitochondrial membrane potential(△·ψm) and Fas/FasL change in lymph,and the effect of dexamethasone on it.Methods The establishment of asthma model .After the inspired collection 2 h,6 h,12 h,24 h,48 h lymph, blood, bronchoalveolar lavage fluid (BALF) of lymphocytes by flow cytometry (FCM) detection of lymphocyte mitochondria transmembrane potential (△·ψm), cell immunohistochemistry (SABC method) was used to detect cell surface Fas FasL protein expression.Results The peak of mitochondrial transmembrane potential in the asthma lymph, blood, BALF lymphocytes was lower compared with the control group and treatment group (P<0.001). There was no significant difference between the control and treatment groups (P>0.05).The Fas, FasL protein optical density on the surface of lymphocytes in the asthma lymph,BALF was obvious lower than that of the control and treatment group, and there was no significant difference between the control group and treatment group.Conclusion The early lymphocyte apoptosis obstacles and the low of Fas / FasL expression were found in asthma rat in lymph,blood and BALF. Particularly, the degree of T lymphocyte apoptosis obstacles and low of Fas / FasL in lymph were remarkable to the blood and BALF in asthma. Dexamethasone may enhance the cell surface through the Fas / FasL expression to accelerate the apoptosis of lymphocytes in asthma.
【Key words】 asthma, lymph,mitochondrial membrane potential,Fas/FasL,apoptosis
支气管哮喘(简称哮喘) 是临床常见的慢性呼吸道疾病,其发病机制尚未明确[1]。诸多研究认为T细胞增多是哮喘炎症持续存在的关键环节[2]。关于哮喘时T细胞增多的机制研究目前大多集中于外周血、肺泡灌洗液(BALF)等组织,而关于哮喘淋巴液免疫状态研究尚未见报道。本文通过检测哮喘大鼠淋巴液中淋巴细胞线粒体跨膜电位、Fas/FasL变化,并观察地塞米松干预的影响,探讨淋巴液中淋巴细胞凋亡与哮喘之间的内在关系。